anti gpnmb primary antibody  (R&D Systems)

Journal: The Journal of pathology

Article Title: GPNMB expression identifies TSC1/2/mTOR-associated and MiT family translocation-driven renal neoplasms.

doi: 10.1002/path.5875

Figure Lengend Snippet: Figure 1. GPNMB is upregulated following TSC2 loss in a TFE3/TFEB- and mTORC1-dependent fashion. (A) Immunoblotting of whole-cell lysates from parental and TSC2 KO HEK293T cells for TSC2 and mTORC1 activation markers. (B) Immunoblotting of whole-cell lysates from parental and TSC2 KO HEK293T cells for GPNMB. A non-specific band (*) based on the siRNA experiment in supplementary material, Figure S2 is present in this cell line by immunoblotting. (C) Immunoblotting of cell lysates enriched for lysosomal content from parental and TSC2 KO HEK293T cells. (D) Immunoblotting of whole-cell lysates from TSC2 KO HEK293T cells with or without 72-h treatment with mTOR inhibitor rapamycin (200 nM). (E) RT-qPCR for relative GPNMB gene expression in TSC2 KO HEK293T cells with or without genomic deletion of TFE3, TFEB, or TFEB and TFE3 (r = 3; ***p < 0.0001 by one-way ANOVA). (F) Immunoblotting of whole-cell lysates from TSC2 KO HEK293T cells with or without genomic deletion of TFE3, TFEB, or TFEB and TFE3. (G) Immunoblotting of whole-cell lysates from TRI-102 parental cells (TSC2/) and TRI-103 cells (TSC2/ with stable transfection of wild-type TSC2) for TSC2 and mTORC1 activation markers. (H) RT-qPCR for relative GPNMB gene expression in TRI-102 and TRI-103 cells. (I) Immunoblotting of cell lysates from TRI102 and TRI103 cells enriched for lysosomal content. (J) Immunoblotting of whole-cell lysates from TRI-102 cells with or without 72-h treatment with mTOR kinase inhibitor Torin 1 (1 μM).

Article Snippet: GPNMB immunohistochemistry (IHC) on human tissues was performed using the Ventana Discovery ULTRA platform (Ventana/Roche, Oro Valley, AZ, USA) using hand-applied GPNMB primary antibody (E4D7P XP® Rabbit mAb #38313, 1:1500; Cell Signaling Technology, Danvers, MA, USA).

Techniques: Western Blot, Activation Assay, Quantitative RT-PCR, Gene Expression, Stable Transfection

Journal: The Journal of pathology

Article Title: GPNMB expression identifies TSC1/2/mTOR-associated and MiT family translocation-driven renal neoplasms.

doi: 10.1002/path.5875

Figure Lengend Snippet: Figure 2. Renal tumors from A/J Tsc2+/ mice show increased GPNMB expression. (A) Hematoxylin and eosin (H&E) and TSC2, p-S6, and p- 4EBP1 IHC staining of cystadenoma lesions occurring in 14-month-old A/J Tsc2+/ mice. Scale bar: 500 μm. (B) Laser capture microdis- section (LCM) of renal cystadenomas from 14-month-old A/J Tsc2+/ mice guided by p-S6 immunostaining for mTORC1 activation. Images reduced from 100 magnification. Scale bar: 250 μm. (C) RT-qPCR for relative GPNMB gene expression in normal kidney parenchyma and LCM-captured renal cystadenoma tissue. (D) H&E and GPNMB IHC staining of oncocytic vacuolated carcinomas occurring in 14-month-old A/J Tsc2+/ mice. Scale bar: 500 μm. Right panels are higher-magnification views of the outlined area in each image.

Article Snippet: GPNMB immunohistochemistry (IHC) on human tissues was performed using the Ventana Discovery ULTRA platform (Ventana/Roche, Oro Valley, AZ, USA) using hand-applied GPNMB primary antibody (E4D7P XP® Rabbit mAb #38313, 1:1500; Cell Signaling Technology, Danvers, MA, USA).

Techniques: Expressing, Immunohistochemistry, Immunostaining, Activation Assay, Quantitative RT-PCR, Gene Expression

Journal: The Journal of pathology

Article Title: GPNMB expression identifies TSC1/2/mTOR-associated and MiT family translocation-driven renal neoplasms.

doi: 10.1002/path.5875

Figure Lengend Snippet: Figure 3. Representative GPNMB immunohistochemical (IHC) staining in common renal cell carcinoma (RCC) subtypes. (A) Representative H&E and GPNMB IHC images for normal renal parenchyma, clear cell RCC (ccRCC), and papillary RCC (papRCC). All images were reduced from 200 magnification. (B) Representative H&E and GPNMB IHC images for oncocytoma and chromophobe RCC (chRCC). Note variable staining in two different chRCC cases. All images were reduced from 200 magnification.

Article Snippet: GPNMB immunohistochemistry (IHC) on human tissues was performed using the Ventana Discovery ULTRA platform (Ventana/Roche, Oro Valley, AZ, USA) using hand-applied GPNMB primary antibody (E4D7P XP® Rabbit mAb #38313, 1:1500; Cell Signaling Technology, Danvers, MA, USA).

Techniques: Immunohistochemical staining, Immunohistochemistry, Staining

Journal: The Journal of pathology

Article Title: GPNMB expression identifies TSC1/2/mTOR-associated and MiT family translocation-driven renal neoplasms.

doi: 10.1002/path.5875

Figure Lengend Snippet: Figure 4. Representative GPNMB staining in translocation RCC (tRCC) cases. Representative H&E and GPNMB IHC images for TFE3-driven tRCC [PRCC–TFE3 and SFPQ–TFE3 cases confirmed by fluorescence in situ hybridization (FISH)] as well as a tRCC negative for cathepsin K immunostaining and a TFEB-driven (FISH-confirmed) tRCC case. All images were reduced from 200 magnification.

Article Snippet: GPNMB immunohistochemistry (IHC) on human tissues was performed using the Ventana Discovery ULTRA platform (Ventana/Roche, Oro Valley, AZ, USA) using hand-applied GPNMB primary antibody (E4D7P XP® Rabbit mAb #38313, 1:1500; Cell Signaling Technology, Danvers, MA, USA).

Techniques: Staining, Translocation Assay, In Situ Hybridization, Immunostaining

Journal: The Journal of pathology

Article Title: GPNMB expression identifies TSC1/2/mTOR-associated and MiT family translocation-driven renal neoplasms.

doi: 10.1002/path.5875

Figure Lengend Snippet: Figure 5. Representative GPNMB immunohistochemical (IHC) staining in TSC1/2/mTOR-related tumors. (A) Representative H&E and GPNMB IHC images for eosinophilic solid and cystic carcinoma (ESC), eosinophilic vacuolated tumor (EVT), and low-grade oncocytic tumor (LOT). All images were reduced from 200 magnification. (B) Representative H&E and GPNMB IHC images for conventional angiomyolipoma (AML) and perivascular epithelioid cell tumor (PEComa). All images were reduced from 200 magnification.

Article Snippet: GPNMB immunohistochemistry (IHC) on human tissues was performed using the Ventana Discovery ULTRA platform (Ventana/Roche, Oro Valley, AZ, USA) using hand-applied GPNMB primary antibody (E4D7P XP® Rabbit mAb #38313, 1:1500; Cell Signaling Technology, Danvers, MA, USA).

Techniques: Immunohistochemical staining, Immunohistochemistry

Journal: The Journal of pathology

Article Title: GPNMB expression identifies TSC1/2/mTOR-associated and MiT family translocation-driven renal neoplasms.

doi: 10.1002/path.5875

Figure Lengend Snippet: Figure 6. Digitally quantified GPNMB expression by renal tumor diagnosis. The point represents the median H-score, and bars repre- sent the interquartile range for each diagnostic group.

Article Snippet: GPNMB immunohistochemistry (IHC) on human tissues was performed using the Ventana Discovery ULTRA platform (Ventana/Roche, Oro Valley, AZ, USA) using hand-applied GPNMB primary antibody (E4D7P XP® Rabbit mAb #38313, 1:1500; Cell Signaling Technology, Danvers, MA, USA).

Techniques: Expressing, Biomarker Discovery, Diagnostic Assay